CD44 KO cell lysate available now. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 1 bp deletion; Frameshift = 98.54%.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 1 bp deletion; Frameshift = 98.54%
Alternative names
BA-1, CD44 antigen, CD44 molecule, CD44 molecule (Indian blood group), CD44_HUMAN, CDW44 antigen, CDw44, CSPG8, Cell surface glycoprotein CD44, ECMR-III, Epican, Extracellular matrix receptor III, GP90 lymphocyte homing/adhesion receptor, HCELL, HSA, HUTCH-I, HUTCH1, Heparan sulfate proteoglycan, Hermes antigen, Hyaluronate receptor, IN, INLU-related p80 Glycoprotein, LHR, MC56, MDU2, MDU3, MGC10468, MIC4, MUTCH I, MUTCH1, PGP-1, Phagocytic glycoprotein 1, Phagocytic glycoprotein I, Soluble CD44, chondroitin sulfate proteoglycan 8, hematopoietic cell E- and L-selectin ligand, homing function and Indian blood group system
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CD44 KO cell lysate available now. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 1 bp deletion; Frameshift = 98.54%.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 1 bp deletion; Frameshift = 98.54%
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- CD44
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
12 product images
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - HRP Anti-CD44 antibody [EPR1013Y] ab194989 observed at 80 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
HRP Anti-CD44 antibody [EPR1013Y] ab194989 was shown to react with CD44 (HRP) in wild-type HeLa cells in western blot Loss of signal was observed when CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938) was used. Wild-type HeLa and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with HRP Anti-CD44 antibody [EPR1013Y] ab194989 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 2500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - HRP Anti-CD44 antibody [EPR1013Y] (HRP Anti-CD44 antibody [EPR1013Y] ab194989) at 1/2500 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 80 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa cell lysate 20 ug
Lane 2: CD44 knockout HeLa cell lysate 20 ug
Lane 3: A549 cell lysate 20 ug
Lane 4: LNCaP cell lysate 20 ug
False colour image of Western blot: Anti-CD44 antibody [C44Mab-5] staining at 1.226 ug/ml, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD44 antibody [C44Mab-5] ab264539 was shown to bind specifically to CD44. A band was observed at 75-80 kDa in wild-type HeLa cell lysates with no signal observed at this size in CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938). To generate this image, wild-type and CD44 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.All lanes: Western blot - Anti-CD44 antibody [C44Mab-5] (Anti-CD44 antibody [C44Mab-5] ab264539) at 1.226 µg/mL
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CD44 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 cell lysate at 20 µg
Lane 4: LNCaP cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 75-80 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa cell lysate 20 ug
Lane 2: CD44 knockout HeLa cell lysate 20 ug
Lane 3: A549 cell lysate 20 ug
Lane 4: LNCaP cell lysate 20 ug
False colour image of Western blot: Anti-CD44 antibody [EPR1013Y] staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD44 antibody [EPR1013Y] ab51037 was shown to bind specifically to CD44. A band was observed at 75-80 kDa in wild-type HeLa cell lysates with no signal observed at this size in CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938). To generate this image, wild-type and CD44 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.All lanes: Western blot - Anti-CD44 antibody [EPR1013Y] (Anti-CD44 antibody [EPR1013Y] ab51037) at 1/5000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CD44 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 cell lysate at 20 µg
Lane 4: LNCaP cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 75-80 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa cell lysate 20 ug
Lane 2: CD44 knockout HeLa cell lysate 20 ug
Lane 3: A549 cell lysate 20 ug
Lane 4: LNCaP cell lysate 20 ug
False colour image of Western blot: Anti-CD44 antibody [EPR18668] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD44 antibody [EPR18668] ab189524 was shown to bind specifically to CD44. A band was observed at 70-85 kDa in wild-type HeLa cell lysates with no signal observed at this size in CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938). To generate this image, wild-type and CD44 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.All lanes: Western blot - Anti-CD44 antibody [EPR18668] (Anti-CD44 antibody [EPR18668] ab189524) at 1/1000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 70-85 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa cell lysate 20 ug
Lane 2: CD44 knockout HeLa cell lysate 20 ug
Lane 3: A549 cell lysate 20 ug
Lane 4: LNCaP cell lysate 20 ug
False colour image of Western blot: Anti-CD44 antibody [SP37] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD44 antibody [SP37] ab101531 was shown to bind specifically to CD44. A band was observed at 75-80 kDa in wild-type HeLa cell lysates with no signal observed at this size in CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938). To generate this image, wild-type and CD44 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.All lanes: Western blot - Anti-CD44 antibody [SP37] (Anti-CD44 antibody [SP37] ab101531) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CD44 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 cell lysate at 20 µg
Lane 4: LNCaP cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 75-80 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 2: Knockout CD44 HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD44 antibody [BLR038F] ab243894 observed at 80 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
Anti-CD44 antibody [BLR038F] ab243894 was shown to react with CD44 in wild-type HeLa cells in western blot Loss of signal was observed when CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938) was used. Wild-type HeLa and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-CD44 antibody [BLR038F] ab243894 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-CD44 antibody [BLR038F] (Anti-CD44 antibody [BLR038F] ab243894) at 1/1000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 80 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD44 antibody [EPR18668] ab189524 observed at 80 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
Anti-CD44 antibody [EPR18668] ab189524 was shown to react with CD44 in wild-type HeLa cells in western blot Loss of signal was observed when CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938) was used. Wild-type HeLa and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-CD44 antibody [EPR18668] ab189524 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-CD44 antibody [EPR18668] (Anti-CD44 antibody [EPR18668] ab189524) at 1/1000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 80 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD44 antibody [EPR1013Y] ab51037 observed at 80 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-CD44 antibody [EPR1013Y] ab51037 was shown to react with CD44 in wild-type HeLa cells in western blot Loss of signal was observed when CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938) was used. Wild-type HeLa and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-CD44 antibody [EPR1013Y] ab51037 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-CD44 antibody [EPR1013Y] (Anti-CD44 antibody [EPR1013Y] ab51037) at 1/5000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 80 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD44 antibody [SP37] ab101531 observed at 80 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-CD44 antibody [SP37] ab101531 was shown to react with CD44 in wild-type HeLa cells in western blot Loss of signal was observed when CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938) was used. Wild-type HeLa and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-CD44 antibody [SP37] ab101531 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-CD44 antibody [SP37] (Anti-CD44 antibody [SP37] ab101531) at 1/1000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 80 kDa
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD44 antibody [MEM-263] ab9524 observed at 80 kDa. Red - loading control, Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
Anti-CD44 antibody [MEM-263] ab9524 was shown to react with CD44 in wild-type HeLa cells in western blot Loss of signal was observed when CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938) was used. Wild-type and CD44 knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-CD44 antibody [MEM-263] ab9524 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 2 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-CD44 antibody [MEM-263] (Anti-CD44 antibody [MEM-263] ab9524) at 2 µg/mL
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 80 kDa
Next Generation Sequencing - Human CD44 knockout HeLa cell lysate (ab263938)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 1 bp deletion; Frameshift = 98.54%
Western blot - Human CD44 knockout HeLa cell lysate (ab263938)
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD44v6 antibody [VFF-7] ab30436 observed at 80 kDa. Red - loading control, Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
Anti-CD44v6 antibody [VFF-7] ab30436 was shown to react with CD44v6 in wild-type HeLa cells in western blot Loss of signal was observed when CD44 knockout cell line Human CD44 knockout HeLa cell line ab262515 (knockout cell lysate ab263938) was used. Wild-type HeLa and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-CD44v6 antibody [VFF-7] ab30436 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-CD44v6 antibody [VFF-7] (Anti-CD44v6 antibody [VFF-7] ab30436) at 1/1000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD44 knockout HeLa cell line (Human CD44 knockout HeLa cell line ab262515)
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 81 kDa, 82 kDa
Observed band size: 80 kDa
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